Rosette Flowering Research Articles

Testing Control Options for Western Salsify (Tragopogon dubius) on Conservation Reserve Program Lands

Western salsify has recently formed dense stands in Conservation Reserve Program (CRP) lands in north-central Montana. Our objective was to test the effects of various herbicide treatments and mowing on western salsify and associated vegetation in CRP lands. Six herbicide treatments and one mowing treatment were applied at three sites in 2010. Herbicide treatments included combinations of glyphosate, 2,4-D, dicamba, and/or metsulfuron-methyl applied when western salsify was either in the rosette or early flowering stage. Mowing was applied at the early flowering stage. Herbicide treatments reduced western salsify and increased perennial grass at one of the three sites, which was the site most dominated by western salsify. When dicamba (0.14 kg ae ha−1) plus 2,4-D (0.48 kg ae ha−1) was applied at the rosette stage, western salsify adult plant density and biomass were reduced to zero and perennial grass biomass increased by 108% in 2010. In 2011, western salsify adult plant density was lower across all herbicide treatments compared to the mowed and nontreated plots. Annual grass density increased by up to 400% when herbicide applications including metsulfuron-methyl were applied at the early flowering stage. Mowing did not control western salsify. Results suggest dicamba plus 2,4-D applied at the rosette stage can provide effective control of western salsify and increase perennial grasses without stimulating the emergence of annual grasses.

Possible contributions of TERMINAL FLOWER 1 to the evolution of rosette flowering in Leavenworthia (Brassicaceae).

Leavenworthia crassa is a rosette flowering species that differs from inflorescence flowering species, such as Arabidopsis thaliana, in having elongated pedicels and shortened interfloral internodes on the main axis. Based on previous experiments, we hypothesized that changes to the L. crassa TFL1 ortholog, LcrTFL1, were important in the evolution of rosette flowering. We isolated LcrTFL1 and introduced a genomic construct into tfl1 mutant A. thaliana plants. We also generated and analyzed EGFP-LcrTFL1 reporter-fusion lines, and LcrTFL1/LcrLFY doubly transgenic lines. The transgene rescued the mutant defects, but manifested gain-of-function phenotypes. However, LcrTFL1 lines differed from 35S:TFL1 lines in several regards. Defects in floral meristem identity establishment were observed, as was the production of flowers with extra petals. We also noted features that resemble rosette flowering: LcrTFL1 lines produced significantly shorter interfloral internodes and significantly longer pedicels than either wild-type or 35S:TFL1 plants. Our data show that there are substantive differences in the regulation and/or function of TFL1 orthologs between A. thaliana and L. crassa. These may reflect changes that occurred during the evolution of rosette flowering in Leavenworthia, but, if so, our results show that additional, as-yet-unidentified genes were involved in this instance of architectural evolution.

Effect of overexpression of Arabidopsis Damaged DNA-binding protein 1A on De-etiolated 1

In Arabidopsis thaliana, de-etiolated 1 mutants (det1) grown in the dark resemble light-grown wild-type seedlings. Arabidopsis DET1 encodes a 62 kD protein, which is a negative regulator of light signaling. UV-damaged DNA-binding protein 1 (DDB1) was initially identified due to its role in human DNA damage repair. Arabidopsis has two DDB1 homologs: DDB1A and DDB1B. DDB1A mutation enhances det1 mutant phenotypes. In this study, we generated Arabidopsis lines that overexpress DDB1A-3HA in wild-type, det1, as well as Myc-DET1 or GFP-DET1 rescued genetic backgrounds. DDB1A-3HA overexpression resulted in decreased apical hook formation in wild-type dark-grown seedlings, and enhanced det1 small rosette and early flowering time phenotypes. In the Myc-DET1 background, DDB1A-3HA overexpression resulted in decreased rescue of dark- and light-grown hypocotyl length, light-grown anthocyanin and chlorophyll levels, adult height and stem number phenotypes. This result is consistent with the decreased levels of Myc-DET1 protein detected in the DDB1A-3HA overexpression line. The GFP-DET1 DDB1A-3HA double overexpression line exhibited increased rescue of dark and light-grown hypocotyl length and light-grown chlorophyll level phenotypes relative to GFP-DET1 alone, despite the fact that GFP-DET1 protein also decreased in the double overexpression line. In addition, increased DET1 resulted in decreased DDB1A-3HA levels due to proteasomal degradation. Overall, DDB1A-3HA overexpression affected phenotypes in a variety of DET1 backgrounds, reduced epitope-tagged DET1 levels, and, correlatively, in general dampened the rescue of det1 mutants by the DET1-DDB1A complex.

Changes in the Essential Oil Content and Chemical Compositions of Echinophora platyloba DC. During Three Different Growth and Developmental Stages

Echinophora platyloba is a valuable essential oil bearing plant. The variation in the quantity and quality of the essential oil of wild populations of E. platyloba extracted by hydro distillation was studied during different growth and developmental stages. The content of essential oils (w/w %) were 0.7, 0.5, and 0.2 % in rosette, floral budding, and full flowering stages, respectively. The essential oils were analyzed by GC and GC-MS. Total of 41, 62, and 56 components were identified and quantified in the above mentioned stages, respectively. The oil content showed remarkable increase at rosette stage. The proportion of (Z)-β-Ocimene, as a major component at rosette, drastically decreased in floral budding and then slightly increased at full flowering stage. In contrast, (E)- β-ocimene was not detected at the rosette stage but was the major component at the subsequent harvest time and then decreased at full flowering stage. Moreover, monoterpene hydrocarbons were the main group of compounds in the rosette (71.6 %), floral budding (64.8 %), and full flowering stage (69.6 %).

An exploration of LEAFY expression in independent evolutionary origins of rosette flowering in Brassicaceae

Whereas most Brassicaceae produce flowers on an elongated inflorescence, a few lineages produce flowers directly from the vegetative rosette on elongated pedicels. Knowing the extent to which independent origins of rosette flowering involve the same developmental and genetic mechanisms could clarify the constraints acting on plant architectural evolution. Prior work in Idahoa, Ionopsidium, and Leavenworthia suggested that changes in the activity or expression of the flower meristem identity gene, LEAFY (LFY), played a role in all three origins of rosette flowering. Here we studied the developmental morphology of L. crassa and immunolocalization of LFY protein in Leavenworthia and Ionopsidium to further compare independent origins of rosette flowering. Leavenworthia crassa differs from Ionopsidium and Idahoa in producing ebracteate flowers. Flowers are, however, associated with "squamules," here interpreted as stipules of a cryptic bract. LFY was detected in L. crassa flower primordia but not in inflorescence meristems. In contrast, the rosette flowering Io. acaule accumulated LFY protein in the inflorescence meristem, whereas its inflorescence-flowering close relative, Io. prolongoi, did not. Thus, although different cases of rosette flowering likely entailed modifications of the same meristem identity program, distinct developmental genetic mechanisms appear to be involved in each case.

The Role of a Floral Identity Gene LFY in Plant Morphological Evolution

The degree to which parallel evolution utilizes the same genetic mechanisms indicates the degree to which developmental processes constrain or channel phenotypic evolution. A transgenetic strategy was used to elucidate the role of one floral meristem identity gene, LEAFY (LFY), in the evolution of rosette flowering, a plant architecture that has evolved in parallel in several lineages of the mustard family, Brassicaceae. The LFY genes from three rosette flowering species were cloned and introduced into a species with the ancestral architecture, and results indicated that changes at the LFY locus contributed to the evolution of rosette flowering in two of the three lineages, but that in each lineage a different set of genetic partners was involved. Also, LFY was shown to play a role in the evolution of flower size. Transgenetic strategy may be useful in the study of plant morphological evolution and parallelism.

Evolutionary divergence of LFY function in the mustards Arabidopsis thaliana and Leavenworthia crassa

LEAFY (LFY), a transcription factor involved in the regulation of flower development in Arabidopsis thaliana, has been identified as a candidate gene in the diversification of plant architecture in Brassicaceae. Previous research with Leavenworthia crassa, which produces solitary flowers in the axils of rosette leaves, has shown that the L. crassa LFY ortholog, LcrLFY, rescues most aspects of flower development in A. thaliana but showed two novel traits: flowers produced additional petals and inflorescences produced terminal flowers. In this paper, we explore the molecular mechanisms responsible for these novel phenotypes. We used microarray hybridizations to identify 32 genes differentially expressed between a transgenic LcrLFY line and a control transgenic LFY line. Of particular interest, TERMINAL FLOWER 1 (TFL1) transcripts were found at elevated levels in LcrLFY lines. To distinguish regulatory versus functional changes within the LcrLFY locus, reciprocal chimeric transgenes between LcrLFY and LFY were constructed. These lines implicate divergence of LcrLFY cis-regulation as the primary cause of both novel transgenic phenotypes but implicate divergence of LcrLFY protein function as the primary cause of elevated TFL1 levels. Taken together these results show that LcrLFY has diverged from A. thaliana in both the cis-regulatory and protein-coding regions and imply that molecular coevolution of LcrLFY and the L. crassa TFL1 ortholog, LcrTFL1, contributed to the evolution of rosette flowering.

Molecular evolution of the transcription factor LEAFY in Brassicaceae

LEAFY ( LFY) is a DNA-binding transcription factor that regulates floral meristem identity. LFY is unusual among angiosperm developmental regulators because it is not part of an extended gene family. Recent expression studies and transgenic experiments have suggested that changes at the LFY locus might have played a role in the evolution of rosette flowering, a modified plant architecture that has evolved at least three times in Brassicaceae. Here we examined the sequences of LFY genes from 16 species of Brassicaceae to evaluate whether gene duplication and/or the shift to rosette flowering correlate with changes in the molecular evolution of LFY. We found evidence of gene duplication in four taxa, but phylogenetic analysis suggested that duplicate genes have generally not persisted through multiple speciation events. This result can be explained if LFY is prone to be lost by drift due to a low probability of subfunctionalization or neofunctionalization. Despite great heterogeneity in dN/dS ratios, duplicate genes show a significant tendency to have elevated dN/dS ratios. Rosette-flowering lineages also show elevated dN/dS ratios and two of the rosette-flowering taxa, Idahoa and Leavenworthia, have some radical amino acid substitutions that are candidates for having played a causal role in the evolution of rosette flowering.

Transgenic study of parallelism in plant morphological evolution.

Developmental constraint is indicated when one finds that similar genetic mechanisms are responsible for independent origins of the same derived phenotype. We studied three independent origins of rosette flowering within the mustard family and attempted to evaluate the extent to which the same mechanisms were involved in each transition from the ancestral phenotype, inflorescence flowering. We used transformation to move a candidate gene, LFY, and its cis-regulatory sequences from rosette-flowering species into an inflorescence-flowering recipient, Arabidopsis thaliana, in place of its endogenous LFY gene. The transgenic phenotypes of experimental and control lines (containing an A. thaliana LFY transgene) and the expression driven by the cis-regulatory sequences show that changes at the LFY locus might have contributed to the evolution of rosette flowering in two of the three lineages. In the third case, changes upstream of LFY are implicated. Our data suggest that changes in a single developmental regulatory program were involved in multiple origins of the same derived trait but that the specific genetic changes were different in each case.

LEAFYand the evolution of rosette flowering in violet cress (Jonopsidium acaule, Brassicaceae)

Arabidopsis and most other Brassicaceae produce an elongated inflorescence of mainly ebracteate flowers. However, the early-flowering species violet cress (Jonopsidium acaule) and a handful of other species produce flowers singly in the axils of rosette leaves. In Arabidopsis the gene LEAFY (LFY) is implicated in both the determination of flower meristem identity and in the suppression of leaves (bracts) that would otherwise subtend the flowers. In this study we examined the role of LFY homologs in the evolution of rosette flowering in violet cress. We cloned two LFY homologs, vcLFY1 and vcLFY2, from violet cress. Their exon sequences show ∼90% nucleotide similarity with Arabidopsis LFY and 99% similarity to each other. We used in situ hybridization to study vcLFY expression in violet cress. The patterns were very similar to LFY in Arabidopsis except for stronger expression in the shoot apical meristem outside of the region of flower meristem initiation. It is possible that the relatively diffuse expression of vcLFY contributes to the lack of bract suppression in violet cress. Additionally, the earliest flowers produced by violet cress express vcLFY, suggesting that accelerated flowering in violet cress could also result from changes in the regulation of vcLFY.

Effects of Temperature and Light Intensity on the Growth of Scrophularia Marilandica

The effects of temperature and light intensity on the growth of Scrophularia marilandica L. were studies under artificial, semi—natural, and natural environmental conditions. The plants in the controlled environment rooms were grown under 10 different combinations of temperature (48°/58°—63°/73°F) and light intensity (24—123 cal/cm2/day). The plants in the semi—natural experiment were grown outdoors on prepared plots representing seven levels of average radiation ranging from 20.4 to 405.7 cal/cm2/day. Net assimilation rate (NAR) was proportional to light intensity up to 123 cal/cm2/day in the growth rooms. In the semi—natural studies, it was proportional up to 150 cal/cm2/day and from this point up to 300 cal/cm2/day, NAR increased but not proportionally. Above 300 cal/cm2/day NAR was independent of solar radiation. Under controlled conditions NAR decreased slightly with age at the highest light levels. The length of the vegetative phase, the specific leaf area, and the leaf—area ratio were sharply decreased at higher light intensities. The dry weight of all plant organs was augmented by high light intensities. Roots and axillary shoots were especially sensitive. In the growth—room experiments, temperature effects were more pronounced than were light effects. Stem elongation was completely inhibited at temperature regimes below 63°/73°F, and leaf expansion was markedly diminished below 57°/68°F. Consequently the shoot developed a rosette habit and flowering was inhibited. Decreases in temperature to 48°/58°F had little effect of root growth. The net effect of low temperatures was to depress the shoot—root ratio. However, NAR was increased by a reduction in temperature from 63°/73° to 48°/58° at all intensities. The shoot—root ratio of shaded plants was increased to a greater extent by a rise in temperature than was that of the unshaded plants. Regardless of light level, plants at 48°/58°F consistently had a smaller shoot—root ratio than did those at 63°/73°F. Inhibition of stem elongation and subsequent rosette development at low temperatures were not affected by changes in light intensity. The usual development of greater leaf areas by plants growing under lower light intensities was partially reversed at low temperatures. In early spring shoots of native Scrophularia marilandica sprouted from crown buds attached to perennial roots. The timing differential between leaf expansion in April and shoot elongation in May appears to be a temperature—photoperiod interaction. In early spring and autumn cool temperatures which repressed shoot growth had little effect on root growth. Roots increased greatly in size through late summer into early autumn.